二维异核 J 分辨核磁共振波谱法定量分析 ¹³C 标记生物样品的同位素特征。

Isotopic profiling of ¹³C-labeled biological samples by two-dimensional heteronuclear J-resolved nuclear magnetic resonance spectroscopy.

机构信息

Université de Toulouse, INSA, UPS, INP, LISBP, F-31077 Toulouse, France.

出版信息

Anal Biochem. 2012 Aug 15;427(2):158-63. doi: 10.1016/j.ab.2012.05.021. Epub 2012 May 29.

DOI:10.1016/j.ab.2012.05.021

Abstract

The use of two-dimensional heteronuclear J-resolved (2D H-JRES) nuclear magnetic resonance (NMR) spectroscopy for fast and reliable measurement of isotopic patterns from ¹³C-enriched compounds resulting from carbon labeling experiments was evaluated. Its use with biological samples of increasing complexity showed that 2D H-JRES spectroscopy is suitable for high-throughput isotopic profiling of any kind of labeled samples. Moreover, the method enabled accurate quantification of ¹³C enrichments and, thus, can be used for metabolic flux analysis. The excellent trade-off between reduced experimental time and the number of measurable isotopic data makes 2D H-JRES NMR a promising approach for high-throughput flux analysis of samples of intermediate complexity.

摘要

二维异核 J 分辨（2D H-JRES）核磁共振（NMR）光谱用于快速可靠地测量碳标记实验产生的 ¹³C 富集化合物的同位素模式，对此进行了评估。随着生物样本复杂性的增加，该方法的使用表明 2D H-JRES 光谱适用于任何类型标记样本的高通量同位素分析。此外，该方法还能够准确量化 ¹³C 丰度，因此可用于代谢通量分析。实验时间和可测量的同位素数据数量之间的出色折衷使得 2D H-JRES NMR 成为高通量分析中等复杂样品通量的有前途的方法。